New 3, 16{60 , 17{60 -trihydroxy-{66 {11 {11 {11 {11 {11 {11 {11 {11 {11 -oestratriene

ABSTRACT

The compound of the formula   Use: Oestrogenic agent for inhibiting the gonadotropic function and ovulation.

United States Patent Anner et al.

[ NEW 3, l6a, l7B-TRIHYDROXY-A mm -OESTRATRIENE [72] Inventors: GeorgAnner; Jaroslav Kalvoda, both of Basel, Switzerland [73] Assignee:Ciba-Geigy Corporation, Ardsley, NY.

[22] Filed: Feb. 11, 1970 [21] Appl. No.: 10,647

511 rm.c| 58 FieldofSearch ..260/397.5

[56] References Cited UNITED STATES PATENTS 3,557,161 1/1971 Dusza eta1. ..260/397.5

[451 July 25, 1972 3,558,776 l/l9 7l Campbelletal ..424/243 PrimaryExaminer-Elbert L. Roberts Attorney-Harry Goldsmith, Joseph G. Kolodnyand Mario A. Monaco I ABSTRACT The compound of the formula HOI Use:Oestrogenic agent for inhibiting the gonadotropic function andovulation.

1 Claim, No Drawings NEW 3, 16a, l7a-TRIHYDROXY-A OESTRA'l-RIENE SUMMARYOF THE INVENTION The new 7a-methyl-'3,l6',17B-trihydroxy-A"-oestratriene processes valuable pharmacological properties. Thus itabove all shows oestrogenic, antigonadotropic, ovulation-inhibitingand/or blastocyteimplantation-inhibiting effects.

The oestrogenic activity can for example be demonstrated in the knownAllen-Doisy test on female rats using doses of 0.003 to 0.3 mg/kg s.c.and of 0.01 to 3 mg/kg p.o., and in the known Biilbring-Burn test, againon female rats, with doses of 0.0003 to 0.3 mg/kg s.c. and 0.003 to 1mg/kg p.o. ln the parabiosis test on rats, the antigonadotropic effectcan furthermore be demonstrated using doses of 0.0003 to 0.03 mg/kg s.c.and 0.01 to 0.3 mg/kg p.o. The ovulation-inhibiting activity can bedemonstrated on normal female rats, using doses of 0.0001 to 0.003 mg/kgs.c. and 0.003 to 0.1 mg/kg p.o. The blastocyteimplantation-inhibitingactivity can be demonstrated on normal rats after copulation, usingdoses of 0.001 to 0.003 mg/kg s.c. and 0.01 to 0.03 mg/kg p.o. The newcompound can thus be used as an oestrogenic agent for inhibiting thegonadotropic function and the ovulation, as well as generally forcontrolling the fertility.

The new compound can be obtained if a compound of formula wherein Rrepresents a free, esterified or etherified hydroxyl group, R representsan oxo group and R represents an esterified hydroxyl group in thea-position together with A hydrogen atom, or R represents a free oresterified hydroxyl group in the B-position together with a hydrogenatom and R represents an oxo group, or R, represents an esterifiedhydroxyl group in the B-position and together with R represents an epoxygroup in the a,a-position, is reduced, the 160:, I713- dihydroxycompound is isolated from the reaction mixture, and if esterified oretherified hydroxyl groups are present in the resulting compound, theformer are split in a manner which is in itself known to give freehydroxyl groups.

The above-mentioned reduction takes place in a manner which is in itselfknown. Thus a l6a-acyloxy-l7-oxo compound or a 16a,l7a-epoxy-l7B-acyloxy compound can be reduced with a di-light metalhydride, preferably lithium aluminum hydride or sodium borohydride. Thisreduction can however also be effected catalytically, for example withhydrogen in the presence of a platinum catalyst. The l6-oxol7B-hydroxycompound, the hydroxyl group of which can also be esterified, ispreferably reduced with nascent hydrogen, such as is for exampleproduced by sodium in an alcohol, or also catalytically in the presenceof, for example, a platinum catalyst. Esterified or etherified hydroxylgroups which may be present are split in the usual manner to give thefree hydroxy group.

An esterified hydroxyl group in the starting substances is preferably ahydroxyl group esterified with a lower aliphatic carboxylic acid, forexample with acetic acid.

The desired compound is separated from the reaction mixture which maycontain isomeric compounds in a manner which is in itself known, forexample by fractional crystallization or chromatography.

The starting substances are known or can be obtained from knowncompounds in a manner which is in itself known. Thus the16,17-epoxy-l7-acyloxy compound can be obtained by converting the known7a-methyl-oestrone into its l7-enolacylate and reaction with a per-acid.The l6a-acyloxy-l7-oxo compound can be obtained therefrom by reactionwith, for example perchloric acid and the l6-oxol 'Ifl-hydroxy compoundcan be obtained therefrom by rearrangement with a base, for examplepotassium carbonate.

The new compound can be used as a medicine in the form of pharmaceuticalpreparation which contain this compound together with pharmaceuticalorganic or inorganic, solid or liquid excipients which are suitable forenteral, for example oral, or parenteral administration. Suitablesubstances for forming these preparations are substances which do notreact with the new compound such as for example water, gelatine,lactose, starch, magnesium stearate, talc, vegetable oils, benzylalcohols, gum, polyalkylene glycols, cholesterol or other knownmedicinal excipients. The pharmaceutical preparations can for example bein the form of tablets, dragees or capsules or in a liquid form assolutions, suspensions or emulsions. They are optionally sterilizedand/or contain auxiliary substances such as preservatives, stabilizers,wetting agents or emulsifiers, salts for regulating the osmotic pressureor buffers. They may also contain further therapeutically valuablesubstances.

The invention is described in more detail in the examples which follow.

EXAMPLE 1 3.0 g of 3,16a-diacetoxy-7a-methyl-l7-oxo-A oestratriene in100 ml of absolute tetrahydrofuran are added dropwise at about 5-10 C.to a suspension of l g of lithium aluminum hydride in 50 ml of absolutetetrahydrofuran, the solution is rinsed down with 100 ml of absolutetetrahydrofuran and the reaction mixture is boiled for 14 hours under areflux condenser. At 5 C. 5 m1 of ethyl acetate in 10 ml oftetrahydrofuran are first added dropwise, followed by 5 ml of wateradded dropwise, the mixture is briefly warmed to about 40 C. andfiltered, and the residue well rinsed with chloroform. The filterresidue is stirred for 1 hour with 300 m1 of 2 N hydrochloric acid atroom temperature, and is filtered off and rinsed with water. The residueis suspended in water and is extracted 4 times with 300 ml at a time ofa chloroformmethanol mixture (4:1). The combined organic phases aredried with sodium sulphate and evaporated in a waterpump vacuum.3,1601,17B-trihydroxy-7a-methyl-A -oestratriene (7a-methyl-oestriol) isthus obtained, which after recrystallization from methylenechloride-methanol-ether melts at 235236 C. [01],?" 55 i 1 (C 0.9]2 inethanol).

The starting material used in this example can for example be obtainedas follows:

About 100 ml are distilled from a solution of 30 g of 7amethyl-oestrone,300 ml of isopropenyl acetate and 19.2 ml of a solution of 40 ml ofisopropenyl acetate and 1.3 ml of concentrated sulphuric acid at normalpressure. After further addition of 300 ml of isopropenyl acetate and19.2 ml of a solution of 40 ml of isopropenyl acetate and 1.3 ml ofconcentrated sulphuric acid about 400 ml are distilled off at normalpressure in the course of a further 3 hours, whereupon, after cooling to5 C., a solution of 42 ml of pyridine in 300 ml of ether is added. Afterdiluting with ice and water the dark mixture is twice extracted with anether-methylene chloride mixture (4:1) and the organic constituents arewashed with water, ice-cold dilute sulphuric acid, water, saturatedsodium hydrogen carbonate solution and again with water until neutral.They are dried over sodium sulphate and evaporated to dryness under awaterpump vacuum. The resulting brown foam is chromatographed on a50-fold quantity of silica gel. On elution with a toluene-ethyl acetatemixture (:5)crude 3,17-diacetoxy-7a-rnethyl-A" '"-oestratetraene isobtained, which after one recrystallization from ether/petroleum ethermelts at 1l0l 11 C. [01],} 77 i 2 0.639).

2.3 g of approximately 88 percent strength m-chloroperbenzoic acid areadded at about 18 C. to a solution of 3.38 g of the resulting compoundin 70 ml of methylene chloride and the mixture is stirred for 30 minutesat room temperature. The

reaction solution is diluted with ether, washed with potassium iodidesolution, sodium thiosulphate solution, water, saturated sodium hydrogencarbonate solution and again with water until neutral, the wash'watersare extracted with ether and the combined organic extracts are driedover sodium sulphate. After evaporation and recrystallization of theresidue'from methylene chloride-ether-petroleum ether 3,l7B-diacetoxy-7a-methyl-l601,17a-oxido-A -oestratriene of melting point 156-157 C. isobtained. [a] 51: 2 (c=0.635).

3.22 g of this compound are dissolved in 32 ml of an aceticaeid-perchloric acid'solution obtained from 49 ml of 96 percent strengthacetic acid and 1 ml of 70 percent strength perchloric acid, and themixture is stirred for 10 minutes at room temperature. The reactionsolution is mixed with ice and water, twice extracted withether-methylene chloride, and the organic extracts are twice washed withwater, dried over sodium sulphate and evaporated to dryness in awater-pump vacuum. The residue is-acetylated overnight with 15 ml ofpyridine and 15 ml of acetic anhydrideThe mixture is then poured ontoice and water, allowed to stand at room temperature, twice extractedwith ether, and the organic phase washed with dilute sulphuric acid,water, saturated sodium hydrogen carbonate solution and again withwater. The extract is dried over sodium sulphate and evaporated under awaterpump vacuum. 3, l 6a-diacetoxy-7a-methyll 7-oxo-A oestratriene isthus obtained, which after recrystallization from ether-petroleum ethermelts at l38-l39 C. [al 135? 1- 2 (c== 0.550).

EXAMPLE 2 A solution of 7.14 g of 3,l7B-diacetoxy-7a-methyl-16a-17a-oxido-A"- "-oestratriene in 285 ml of tetrahydrofuran is added to asuspension of 2.85 g of lithium aluminum hydride in 285 ml oftetrahydrofuran at about l0l5 C. After rinsing down with 140 ml oftetrahydrofuran, the reaction mixture is boiled for 2 hours under areflux condenser, treated at about 10 C. firstly with 30 ml of ethylacetate and then with 550 ml of 2N hydrochloric acid, and 1 liter ofchloroform is added. The mixture is stirred for 10 minutes at roomtemperature, and the organic constituents are separated off, washed withwater, dried over sodium sulphate and evaporated to dryness in awaterpump vacuum. The residue is adsorbed on a 50-fold amount of silicagel and the acid is eluted with a 7:3 mixture of toluene 'and ethylacetate and then with ethyl acetate.

Evaporation of the eluate obtained with ethyl acetate yields 3, 16a, 17fl-trihydroxy-7a-methyl-A -oestratriene, which afier recrystallizationfrom methylene chloride/methanol/ether melts at 235-236C. [01],, +55 l(c=0.9l2 in ethanol).

EXAMPLE 3 Six-hundred mg of 3,17a dihydroxy-7a-methyl-l6-oxo-A--=""-oestratriene are refluxed in 50 ml of isopropanol. In the courseof 20 minutes, 2.0 g of sodium are added in portions.

When all of the sodium has dissolved, the cooled solution is poured intoa mixture of ice and water, and the batch is acidified with dilutehydrochloric acid. The substance which precipitates is filtered off anddissolved in methylene chloridehnethanol, the solution dried over sodiumsulfateand evaporated in a water-jet vacuum, then chromatographed overthe 30-fold quantity of silica gel. Elution with a 30:70 mixture oftoluene and ethyl acetate yields 365 mg of crude 7a-methyl estriol whichafter recrystallization from methylene chloride-i-methanoH-235-236 C.

The starting material used in this example can be prepared for exampleas follows:

4.0 g of solid 3,l6a-diacetoxy-7a-methyl-l7-oxo-A-- estratriene areadded to a briefly boiled suspension of 5.0 g of potassium carbonate in100 ml of methyl alcohol. The reaction solution is then refluxed forminutes under nitrogen, then poured into a mixture of ice and water,acidified with dilute hydrochloric acid, and filtered. The filter cakeis dried and dissolved in methylene chloride, and filtered through g ofalumina (activity stage II, neutral) to eliminate a polar contamination.2.50 g of pure, non-crystallizing 3,17B-dihydroxy- 7a-methyll 6-oxoA"-oestratriene are obtained.

We claim:

